1. Bacterial Biotechnology

2. The Structure of Microbes Prokaryotes Archaebacteria Includes halophiles, thermophiles, “extremophiles” Eubacteria On skin, pathogens, soil, water Generally smaller than Eukaryotes (1-5μm vs 10-100 μm) What are some other characteristics of prokaryotes? (cell wall (gram stain), no nucleus, binary fission, 20 min growth rate…) Do you know how to isolate single colonies?

3. Microorganisms as Tools Microbial Enzymes Taq (DNA polymerase), cellulases, proteases, amylases Bacterial Transformation The ability of bacteria to take in DNA from their surrounding environment Cells must be made competent (to take up DNA)

5. Electroporation A mixture of bacteria and plasmid are briefly electrically shocked

7. Cloning and Expression Techniques Fusion Proteins Use recombinant DNA methods to insert the gene for a protein of interest into a plasmid containing a gene for a well-known protein that serves as a “tag” The tag allows for isolation and purification Ex. GFP – Green Flourescent Protien Used to follow certain molecules thorugh cell processes

9. Microbial Proteins as Reporters Examples: the lux gene which produces luciferase Used to develop a fluorescent bioassay to test for TB (the lux gene is in a virus that only infects M. tuberculosis). If the bacteria is present, the virus infects the cells and the bacterial cells glow!

10. Using Microbes for a Variety of Everyday Applications Food Products Rennin used to make curds (solid) and whey in production of cheese Recombinant rennin is known as chymosin (first recombinant food product approved by FDA)

11. Therapeutic proteins Recombinant insulin in bacteria What is Type I diabetes (insulin-dependent diabetes mellitus) Inadequate production of insulin by beta cells in the pancreas

13. Field Applications of Recombinant Microorganisms Ice-minus bacteria (remove ice protein producing genes from P. syringae) P. fluorescens containing the gene that codes for the bacterial toxin from Bacillus thuringiensis (kills insects) Bt toxin!

14. Using Microbes Against Other Microbes Antibiotics Penicillin was the first Act in a few key ways Prevent replication Kill directly Damage cell wall or prevent its synthesis How do antibiotic resistant strains arise? How can studying bacterial pathogens lead to new drugs?

16. Microbial Genomes Microbial Genome Program (MGP) –the goal is to sequence the entire genomes of microorganisms that have potential applications in environmental, biology, research, industry, and health Sequencing Strategies 

18. Microbial Diagnostics Using Molecular Techniques to Identify Bacteria RFLP - restriction fragment length polymorphism PCR – Polymerase Chain Reaction PulseNet (Contaminated food) - a network run by the Centers for Disease Control and Prevention (CDC) which brings together public health and food regulatory agency laboratories around the United States Centers for Disease Control and Preventionpublic healthUnited States

20. Combating Bioterrorism The use of biological materials as weapons to harm humans or animals and plants we depend on for food Examples in History Throwing plague infected dead bodies over the walls of their enemies Using Biotech Against Bioweapons Postal service x-raying packages Antibody tests in the field PCR tests in the field

21. RISE OF THE SUPERB UGS! An intro to bacteria, infectious diseases, and antibiotic resistance

22. RISE OF THE SUPER BUGS! An intro to bacteria, infectious diseases, and antibiotic resistance

23. How abundant are bacteria? Bacterial Abundance Total bacteria on Earth5 x 10 30 Number of stars in the universe7 x 10 22 Age of the universe in seconds4.4 x 10 17 Bacteria in the human gut1 x 10 14 Global gross product ($/year)7 x 10 13 Cells in the human body1 x 10 13 Texts sent in 20091.5 x 10 12 People on Earth6.9 x 10 9 10X

24. How many types of bacteria are there? Bacterial Distribution Insect Species1-10 million Bacterial species in the soil4 million Bacterial species in the air4 million Bacterial species in the ocean2 million Bird Species10,000 Bacterial species in the human mouth600 Bacterial Species in the human gut500 Pathogenic Species55

25. Infectious Diseases Pathogen: a biological agent that causes disease Pathogen: a biological agent that causes disease Can be a virus, bacteria, fungi, protozoa, or parasite Can be a virus, bacteria, fungi, protozoa, or parasite Infectious diseases are communicable or transmissible from one person to another Infectious diseases are communicable or transmissible from one person to another

26. Virus: Influenza (H1N1) Fungi: Plant pathogen (Rigidoporus laetus) Protist: Malaria (Plasmodium)

27. Mycobacterium tuberculosisSalmonella typhimurium Staph Infection: Staphylococcus aureus

28. Antibiotics Compounds that kill bacteria or inhibit their growth Compounds that kill bacteria or inhibit their growth Naturally produced by microorganisms to kill others Naturally produced by microorganisms to kill others Now many are made synthetically Now many are made synthetically

29. Why is resistance bad? Because you can’t treat the disease! Because you can’t treat the disease! The patient will remain sick  The patient will remain sick 

30. How do you get resistance? Usually from mutation in DNA Usually from mutation in DNA Genetic mutations can be passed to offspring… Genetic mutations can be passed to offspring… Leads to evolution! Leads to evolution!

31. Genetic Mutation NATURAL SELECTION!

32. Susceptible + Antibiotic Antibiotic Resistant

33. What is MRSA?

34. MRSA MRSA stands for Methicillin-Resistant Staphylococcus aureus MRSA are Staph aureus bacteria that have become resistant to this antibiotic.

35. In general, healthy people are at low risk of getting sick with MRSA. Some risk factors include: Recurrent skin diseases or open wounds Long-term illness or long-term dialysis patient Illicit injecting drug use Surgery

36. Been a patient in the hospital or other health care facility within the past year Contact with other persons with MRSA infection Recent antibiotic use Live in crowded settings

37. HOW ARE THESE GERMS SPREAD? MRSA is transmitted primarily by contact with a person who has an infection or is colonized with the bacteria.

38. And now, let’s look at what MRSA really looks like…

41. NDM-1 New Delhi Metallo-beta-lactamase-1 (NDM-1) an enzyme that makes bacteria resistant to a broad range of beta-lactam antibioticsenzymebacteriaresistantbeta-lactam antibiotics first detected in a Klebsiella pneumoniae isolate from a Swedish patient of Indian origin in 2008Klebsiella pneumoniaeSwedishIndian Later detected in India, Pakistan, the United Kingdom, the United States, Canada, and JapanIndiaPakistanUnited KingdomUnited StatesCanada

42. Usually found in: Escherichia coli and Klebsiella pneumoniae, Escherichia coli but the gene for NDM-1 can spread from one strain of bacteria to another by horizontal gene transfer!horizontal gene transfer

43. Acknowledgements The Infectious Disease Control Unit of the Department of State Health Services thanks Ginger Shields, RN Emergency Preparedness Specialist Texarkana-Bowie County Health Department for the creation and use of this presentation.